A Study on Isolation, Purification, and Microencapsulation of L-Asparaginase From Soil Bacteria

Author(s): Pooja Sahu and S.T. Asheeba

Abstract: L-asparaginase is considered to be a therapeutic enzyme, which is used to treat acute lymphoblastic leukaemia. Soil serves as potential source of bacteria, which can produce L-asparaginase enzyme. In the present study, L-asparaginase enzyme producing bacteria was isolated from soil, using serial dilution technique on modified M-9 media and screened for the production of L-asparaginase enzyme. Out of 6 bacterial strain one pink coloured bacterial strain were subcultured on modified M-9 broth. Morphological and biochemical analysis was done for bacterial identification and confirmed through 16S rRNA analysis as Enterobacter cloacae. The isolated bacterial strain was further subjected to enzyme production and purification. The activity of the enzyme after ammonium sulphate precipitation 0.025 unit/mg, dialysis 0.015 unit/mg, and ion-exchange chromatography 0.013 unit/mg was recorded. The molecular weight of the finally purified enzyme was found to be 35 kDa. Further the purified enzyme was encapsulated in sodium alginate microsphere and size ranged in 1 mm. In vitro release study was done and the rate of release of encapsulated enzyme was recorded by estimating of enzyme activity as 0.039 unit/mg.

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